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Open AccessCase study Effect of pioglitazone treatment on behavioral symptoms in autistic children Marvin Boris*1, Claudia C Kaiser2, Allan Goldblatt1, Michael W Elice1, Stephen M Edels

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Open Access

Case study

Effect of pioglitazone treatment on behavioral symptoms in autistic children

Marvin Boris*1, Claudia C Kaiser2, Allan Goldblatt1, Michael W Elice1,

Stephen M Edelson3, James B Adams4 and Douglas L Feinstein2

Address: 1 77 Froehlich Farm Blvd Woodbury, New York 11797, USA, 2 Department of Anesthesiology, University of Illinois, Chicago, IL, 60612, USA, 3 Autism Research Institute, 4182 Adams Ave, San Diego, CA 92116, USA and 4 Arizona State University, PO Box 876006, Tempe, AZ

85287-6006, USA

Email: Marvin Boris* - mboris@pol.net; Claudia C Kaiser - ckaiser@uic.edu; Allan Goldblatt - mboris@pol.net;

Michael W Elice - mboris@pol.net; Stephen M Edelson - edelson4@comcast.net; James B Adams - jim.adams@asu.edu;

Douglas L Feinstein - dlfeins@uic.edu

* Corresponding author

Abstract

Introduction: Autism is complex neuro-developmental disorder which has a symptomatic

diagnosis in patients characterized by disorders in language/communication, behavior, and social

interactions The exact causes for autism are largely unknown, but is has been speculated that

immune and inflammatory responses, particularly those of Th2 type, may be involved

Thiazolidinediones (TZDs) are agonists of the peroxisome proliferator activated receptor gamma

(PPARγ), a nuclear hormone receptor which modulates insulin sensitivity, and have been shown to

induce apoptosis in activated T-lymphocytes and exert anti-inflammatory effects in glial cells The

TZD pioglitazone (Actos) is an FDA-approved PPARγ agonist used to treat type 2 diabetes, with a

good safety profile, currently being tested in clinical trials of other neurological diseases including

AD and MS We therefore tested the safety and therapeutic potential of oral pioglitazone in a small

cohort of children with diagnosed autism

Case description: The rationale and risks of taking pioglitazone were explained to the parents,

consent was obtained, and treatment was initiated at either 30 or 60 mg per day p.o A total of 25

children (average age 7.9 ± 0.7 year old) were enrolled Safety was assessed by measurements of

metabolic profiles and blood pressure; effects on behavioral symptoms were assessed by the

Aberrant Behavior Checklist (ABC), which measures hyperactivity, inappropriate speech,

irritability, lethargy, and stereotypy, done at baseline and after 3–4 months of treatment

Discussion and evaluation: In a small cohort of autistic children, daily treatment with 30 or 60

mg p.o pioglitazone for 3–4 months induced apparent clinical improvement without adverse

events There were no adverse effects noted and behavioral measurements revealed a significant

decrease in 4 out of 5 subcategories (irritability, lethargy, stereotypy, and hyperactivity) Improved

behaviors were inversely correlated with patient age, indicating stronger effects on the younger

patients

Conclusion: Pioglitazone should be considered for further testing of therapeutic potential in

autistic patients

Published: 05 January 2007

Journal of Neuroinflammation 2007, 4:3 doi:10.1186/1742-2094-4-3

Received: 13 November 2006 Accepted: 05 January 2007 This article is available from: http://www.jneuroinflammation.com/content/4/1/3

© 2007 Boris et al; licensee BioMed Central Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Autism, the most common of the group of disorders

col-lectively referred to as Autism Spectrum Disorders (ASD),

is a complex neurological disease of unknown etiology

The incidence of autism is estimated to be 1 per 166 [1]

with a male to female ratio of 4:1 Autism has been found

throughout the world in families of all racial, ethnic and

social backgrounds Although accumulating evidence

sug-gests that genetic, environmental, inflammatory,

immu-nological, and metabolic factors play a prominent role in

this disease [2-7], the precise causes remain to be

deter-mined

Altered immune responses in children with ASD are well

documented Autoimmune disorders of thyroiditis,

coli-tis, myelin basic protein autoantibodies, and diabetes are

prevalent in children with ASD Stubbs (1976) published

that 5 of 13 autistic children had no detectable rubella

antibodies despite prior immunization [7] An additional

study showed peripheral mononuclear cells had a

decreased proliferative response to mitogenic stimulation

compared to normal children [8] These findings of

abnormal T-lymphocyte function have been replicated by

other investigators [9,10] Inflammatory responses in

ASD have also been reported to occur in brain, for

exam-ple neuroinflammatory processes involving both

micro-glia and astromicro-glia were found on post mortem

examination in autistic children with elevated cytokine

levels in the cerebral spinal fluid [11,12] Children with

ASD have increased cytokines of Th2 and Th1 arms of the

immune response with Th2 predominant without an

increase in IL10 [13]

Peroxisome proliferator-activated receptor gamma

(PPARγ) is a nuclear hormone receptor originally

charac-terized by its ability to regulate adipocyte differentiation

and gene transcription [14] PPARγ agonists include fatty

acids, non-steroidal anti-inflammatory drugs (NSAIDs),

the natural compound 15-deoxy12,14-prostaglandin-J2

(PGJ2), and members of the class of synthetic drugs

termed thiazolidinediones (TZDs) which include

piogli-tazone (Actos) and rosiglipiogli-tazone (Avandia) TZDs were

originally designed as anti-diabetic drugs due to their

insulin sensitizing effects, and several are now in clinical

use In addition to insulin sensitizing effects, TZDs also

exert anti-inflammatory effects on a variety of cell types,

and for this reason some are being considered for

treat-ment of inflammatory diseases including artherosclerosis

[15], psoriasis [16,17], and inflammatory bowel disease

[18-21] TZDs also reduce inflammatory activation of

brain glial cells, and increase metabolic activities in glial

cells which can lead to increased glucose uptake, lactate

production, and mitochondrial function [22,23]

Further-more, pioglitazone can cross the BBB, [24] suggesting

pos-sible direct effects on brain physiology, which could

positively influence possible abnormalities in regional brain glucose utilization [25] or dysregulation of func-tional activity [26] as reported to occur ASD

The safety and efficacy of pioglitazone has been estab-lished by clinical studies worldwide [27,28] and since FDA approval, pioglitazone has been prescribed to several million patients The adverse events associated with TZDs including pioglitazone are generally mild and transient, and those effects returned to baseline upon withdrawal from, or completion of the studies Two recent studies for the treatment of diabetes in adolescents point to a good safety profile for Actos in younger populations [29,30] Studies with PPARγ drugs in animal models of neurologi-cal conditions have led to clinineurologi-cal testing of these drugs in Alzheimer's disease (AD) and multiple sclerosis (MS) [31,32] These properties of PPARγ agonists make them promising candidates for a therapeutic approach to influ-ence the clinical course of ASD In this report we discuss initial findings using pioglitazone to treat children with autism, which provides the rationale for design of larger clinical trials

Case description

Population

The autistic children all were patients of Marvin Boris,

MD, Allan Goldblatt, PA, and Michael Elice, MD Twenty-five children and adolescents participated in this study The mean age was 7.9 ± 3.5 years, with a range from 3 to

17 years There were 22 males and 3 females All of the participants received an independent diagnosis of Autism Spectrums Disorder (ASD) from an independent clinician and/or agency None of the children had diagnosed Asperger's Disorder (a mild variant of ASD with higher social functioning) or PDDNOS (Pervasive Developmen-tal Disorder – Not Otherwise Specified, a condition with social or behavioral impairments but which do not meet the DSM-IV criteria for ASD) The diagnosis of autism was initially established by a board certified pediatric neurol-ogist, developmental pediatrician, or psychiatrist with experience in ASD In addition, at the first visit to the offices of the treating physician, the child had to meet the DSM-IV checklist criteria for ASD All the children had been receiving behavioral and educational therapies These included speech, occupational, and physical ther-apy, applied behavioral analysis, and auditory integration therapy The children had also received various biomedi-cal interventions for at least one year These included dairy and gluten free diet, metabolic treatment with supple-ments to known deficiencies such as MTHFR (methylene-tetrahydrofolate reductase), treatment with intravenous gamma globulin or secretin, vitamin supplementation, and heavy metal chelation The children who responded poorly (no noticeable improvements in cognitive, social, behavior, or language skills) for at least one year to

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bio-medical, behavioral, or educational therapies were

selected to be treated with pioglitazone as part of the

rou-tine health care treatment, based on papers suggesting

that ASD includes an auto-immune or inflammatory

com-ponent [33,34], and that pioglitazone can reduce T-cell

activation and Th2-type cytokine production, both

impli-cated in ASD [35-38] The rationale and risks of taking

pioglitazone were explained to the parents, and parental

written consents were obtained for all participants A

ret-rospective review of their personal medical records was

approved by the Internal Review Board of Arizona State

University

Comorbities

The autistic population has well-known auto-immunne

comorbidities In this group of autistic children, 7/25

(28%) had thyroiditis, 8/25 (32%) had colitis, 8/25

(32%) had PANDAS (Pediatric acquired neurological

dis-order associated with streptococcus), 20/25 (80%) had

allergic diseases, and 7/25 (28%) were positive for serum

antibodies to myelin basic protein In addition 2/25 had

seizures prior to being treated with pioglitazone

Treatment

Children were prescribed pioglitazone either 30 mg per

day, p.o for ages 3–5 years old; or 60 mg per day for ages

6–17 years old These children were followed with

monthly complete blood counts, glucose and insulin

lev-els, and serum metabolic assays

Analysis

The participants' parents completed the Aberrant

Behav-ior Checklist (ABC) prBehav-ior to the administration of

piogli-tazone and then at a follow-up assessment, 12 or 16 weeks

later There are five subscales on the ABC, consisting of 58

questions The subscales are: hyperactivity, inappropriate

speech, irritability, lethargy, and stereotypy Each

ques-tion was rated on a 4-point scale: 0 = 'not a problem,' 1 =

'the behavior is a problem but slight in degree,' 2 = 'the

problem is moderately serious,' and 3 = 'the problem is

severe in degree.' 'The ABC has been shown to be a valid

and reliable procedure to evaluate treatment efficacy

[39-41] Each of the five subscales was analyzed using paired

t-tests The relationship between age and amount of

behavior change was examined using Pearson product

correlations

Outcomes

There were no significant abnormalities observed in

standard blood analyses in the group of 25 autistic

chil-dren treated with pioglitazone for up to 4 months (Table

1) Over the course of treatment, there were no elevations

in hemoglobin, creatine, BUN (blood urea nitrogen) or

insulin levels There were 2 incidents of slightly and

tran-siently elevated white blood counts and glucose levels,

and 3 incidents of slightly and transiently elevated liver enzyme (ALT and AST) levels All elevations resolved without interventions

A comparison of the mean scores for ABC subscales between baseline and end of treatment for each of the patients revealed that four of the five ABC subscales decreased significantly following the administration of pioglitazone (Figure 1) These subscales were hyperactiv-ity, irritabilhyperactiv-ity, lethargy, and stereotypy There was no change in inappropriate speech; however, it should be noted that the speech subscale is of limited value in chil-dren with autism who lack or have very limited speech

Of the 25 patients, 76% showed an improvement (defined as >50% decrease in score) in at least one sub-group; while 56% showed an improvement in two or more subgroups, and 40% showed improvements in 3 or more subcategories If response rate is estimated as those who showed >25% decrease in at least 2 of the 5 sub-scales, then the percentage is much higher 71% The majority of patients (52%) showed an improvement (>50%) in the hyperactivity subscale

Significant inverse correlations (Figure 2) were detected between age and the improvements calculated for irrita-bility (P = 0.03), lethargy (P = 0.02) and hyperactivity (P

= 0.007) This indicates a tendency for younger partici-pants to benefit more from pioglitazone than the older participants

Discussion and evaluation

The current study provides evidence that treatment with the PPARγ agonist pioglitazone (Actos) does not induce any significant adverse effects, and may have a beneficial effect on patterns of aberrant social behavior in children with diagnosed autism Despite the small sample size (n

= 25 total), we observed statistically significant decreases

in 4 of the 5 subscales of the ABC after a relatively short (4 months) treatment with pioglitazone It is yet not known

Table 1: Incidents of elevated blood values

1 White blood cell counts, normal range 3.8 to 10.5 × 1000 cells per mcl Values of 11.0 and 12.0 recorded 2 Glucose, normal range 70–99 mg/dl Values of 102 and 106 recorded 3 Aspartate aminotransferase, normal range 10–40 IU/L Values of 42, 48, and 45 recorded 4 Alanine aminotransferase, normal range 10–45 IU/L Values of 56, 60, and 48 recorded 5 ALT and AST elevations occurred in the same three patients 6 Pre, pre-trial; Mid, mid-trial; Post, post-trial.

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if these improvements are long lasting, or if they will

con-tinue after treatment is withdrawn Although originally

approved for treatment of Type 2 diabetes in adults, recent

clinical trials of pioglitazone for treatment of diabetes in

adolescents suggest this drug will be well tolerated in

younger populations [29,30]

There is increasing evidence for an association of ASD

with various immune syndromes It was reported that

66% of children with autism have a relative with an

autoimmune disease [42], and families of children with

PDD (Pervasive Development Disorder) have a higher

average number of autoimmune diseases than families of

healthy children [43] Recently the occurrence of AITD

(Autoimmune Thyroid Disease) in first or second order

relatives was concluded to be a risk factor for those ASD

children who show regression (the early loss of already

established skills of communication or of social

interac-tions) [44] The possibility therefore exists that

pioglita-zone influences some aspect of auto-immune nature in

ASD children

It has been suggested that a Th2-like dysfunction may

con-tribute to the causes of ASD In children with ASD, a

pre-ponderance of Th2-like (IL4, IL6, IL10) over Th1-like (IL2,

IFNg, IL1β) cytokines has been reported [45-48] These

studies support the idea that a predominance of Th2

cytokines may be a factor in ASD PPARγ agonists are

known to influence T-cell physiology, and although most

often they have been shown to reduce Th1-like cytokine

(IL1β, TNFa, IL12) production, in several studies they also

reduced Th2 responses In CD4 cells, PGJ2 and the TZD

ciglitazone reduced IL4 production [35] and in EAE, the

animal model of Multiple Sclerosis, PGJ2 blocked splenic

T cell production of IL10 and IL4 [36] PPARγ agonists

also reduce the clinical symptoms in animal models of asthma, a disease which is also thought to be predomi-nantly Th2 type involving IL4, IL5, and IL13 [37] PPARγ agonists have been shown to reduce IL4, IL5, and IL13 production from Tcells of mice with induced lung inflam-mation [38,49] However, in one study the TZDs increased IL4 and IL10, and stimulated GATA3 expression (a transcription factor which shifts cells towards Th2

phe-Relationship of behavioral improvements to age

Figure 2 Relationship of behavioral improvements to age

Dif-ferences in scores for the 5 subscales of the ABC were calcu-lated and plotted versus patient age, and analyzed using Graphpad Prism V4 assuming Gaussian distributions

Effect of Pioglitazone on behavior improvement

Figure 1

Effect of Pioglitazone on behavior improvement The

average (mean ± s.d.) of the total scores for the 5 subscales

of the ABC was calculated for 25 patients before treatment

(baseline) and after 3–4 months of treatment with

Pioglita-zone *, P < 05 unpaired T-test

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notype) [50]; although in other studies PPARγ drugs were

shown to inhibit GATA3 activity [51,52] Nevertheless,

taken together these studies demonstrate that PPARγ

ago-nists have the potential to shift the T-cell response from

Th2 to Th1, or to reduce Th2 cytokine expression, which

may be of therapeutic benefit in ASD

Despite observing significant improvements in 4 of 5

sub-scales of the ABC, the open-label nature of this study

lim-its the ability to draw strong conclusions regarding

treatment-dependent benefits In addition, well-known

expectancy effects in the parent population make

interpre-tation of the ABC subject to potential bias [53,54] The

placebo effect in ASD has been reported to be high in

some studies where improvement was assessed using the

ABC Improvements occurred in 25% of patients

follow-ing atomoxetine treatment for 6 weeks, [55]; 34% after 8

week treatment with risperidone [56]; and 37% after 3

weeks treatment with amantadine [54] In the current

study, the number of responders (those showing >50%

improvement in at least one subscale) was 76%,

consider-ably higher than the values reported in the above studies

An additional confound of the current study is the

diver-sity of auto-immune comorbidities that are common in

the autistic population It is possible that pioglitazone

effects are, in part or in full, an indirect consequence of

reducing symptoms of the autoimmune diseases present

in the study population (thyroiditis, colitis, and

PAN-DAS) For example, in autoimmune thyroiditis (AITD),

pioglitazone could increase levels of suppressor T-cells

that are deficient [57] and as a result reduce circulating

levels of Th1 or Th2 cytokines Similarly, activation of

PPARγ can suppress experimentally induced colitis [58]

which could also reduce plasma cytokine levels, and in

fact several clinical trials of PPARγ agonists for treating

colitis are in progress [19,59] PANDAS, a pediatric

autoimmune neuropsychiatric disorder associated with

streptococcal infections is defined by

obsessive-compul-sive (OCD) and or tic disorders, is thought to be due to

the actions of auto-immune antibodies on basal ganglia

neurons [60], and is improved by immunomodulatory

therapies [61]; anti-inflammatory effects of PPARγ

ago-nists could therefore influence the course of this disease

However, since the precise relationships between

autoim-mune diseases and the penetrance of autistic symptoms

remains to be established, deciphering the relative

impor-tance of indirect effect of pioglitazone on behavior will be

a formidable task

The recent increase in type 2 diabetes in children has

resulted in an increased interest of researchers to explore

the use of anti-diabetic drugs including TZDs in children,

therefore providing additional information regarding the

safety of TZDs in this population A recent clinical trial

tested the effects of rosiglitazone (2 mg bid increased to 4

mg bid after 8 weeks), a related TZD, in 195 obese type 2 diabetic children (age range 8–17 years), in a 24-week double-blind, randomized, metformin-controlled, paral-lel group design The rosiglitazone group gained ~3 kg after 24 weeks with the occurrence of peripheral edema in

1 child [29] However, no other adverse effects were reported, suggesting that TZDs are well tolerated in chil-dren as in adults More recently [30] pioglitazone (15 mg

po escalated to 30 mg po after 4 weeks) was tested as an adjunct therapy for the treatment of type 1 diabetes in a small group of young adolescents (age range 10–17.9 years) After 6 months treatment the pioglitazone subjects showed a small but significant increase in BMI z-score (body mass index standard deviation for age) suggesting treatment-related weight gain In the 35 subjects who completed the study, there was no evidence of edema, anemia, or of any significant increase in the frequency of hypoglycemia in the treatment group versus the placebo group However, it is clear that the safety of pioglitazone, and of other TZDs, in the pediatric population requires additional testing

Conclusion

In view of its established safety profile, the current results provide the rationale for further testing of pioglitazone in autism and other forms of ASD

Abbreviations

ABC: Aberrant Behavior Checklist AD: Alzheimer's disease

ASD: Autism Spectrum Disorder BBB: Blood brain barrier CBC: Complete blood count CD: Cluster of differentiation IL: Interleukin

MS: Multiple Sclerosis NSAID: Non steroidal anti-inflammatory drug PANDAS: Pediatric autoimmune neuropsychiatric disor-der associated with streptococcal infections

PGJ2: 15-deoxy-delta12,14-prostaglandin J2 PDD: pervasive developmental disorder PPAR: Peroxisome proliferator activated receptor

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TNF: Tumor necrosis factor

TZD: thiazolidinedione

Competing interests

The author(s) declare that they have no competing

inter-ests

Authors' contributions

MB and AG were the primary physicians who treated the

patients, and carried out behavioral testing to determine if

the medication was helping their patients CK prepared

the first draft of the paper, and analyzed the data DLF

organized and analyzed the data, contributed to the

orig-inal idea to treat ASD patients, helped write and edit the

manuscript

Acknowledgements

The authors wish to acknowledge the financial assistance of the Autism

Research Institute (San Diego, CA) and dedicate this study to the memory

of its founder Dr Bernard Rimland,

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